U.S. patent application number 10/399701 was filed with the patent office on 2004-02-05 for recombined anti-gpiib/iiia antibodies used for inhibiting angiogenesis.
Invention is credited to Berchtold, Peter, Escher, Robert.
Application Number | 20040022791 10/399701 |
Document ID | / |
Family ID | 7663917 |
Filed Date | 2004-02-05 |
United States Patent
Application |
20040022791 |
Kind Code |
A1 |
Berchtold, Peter ; et
al. |
February 5, 2004 |
Recombined anti-gpiib/iiia antibodies used for inhibiting
angiogenesis
Abstract
The present invention relates to the use of special
phage-display-optimized antibodies directed against GPIIb/IIIa for
jointly inhibiting fibrinogen binding to platelets and vitronectin
binding to endothelial cells for the purpose of the therapy and/or
prophylaxis of vascular occlusion. The present invention
furthermore relates to the use of the antibodies for inhibiting
angiogenesis and/or for inhibiting the metastasis of tumors and/or
for inhibiting intima hyperplasia following vascular damage. bmmy
17.11.2000
Inventors: |
Berchtold, Peter;
(Hinterkappeln, CH) ; Escher, Robert; (Berne,
CH) |
Correspondence
Address: |
ROTHWELL, FIGG, ERNST & MANBECK, P.C.
1425 K STREET, N.W.
SUITE 800
WASHINGTON
DC
20005
US
|
Family ID: |
7663917 |
Appl. No.: |
10/399701 |
Filed: |
April 21, 2003 |
PCT Filed: |
November 20, 2001 |
PCT NO: |
PCT/EP01/13445 |
Current U.S.
Class: |
424/155.1 |
Current CPC
Class: |
C07K 16/2848 20130101;
C07K 2317/50 20130101; A61P 9/10 20180101; A61P 43/00 20180101;
A61P 9/00 20180101; A61P 7/02 20180101; C07K 2317/565 20130101;
A61P 35/04 20180101; A61K 2039/505 20130101 |
Class at
Publication: |
424/155.1 |
International
Class: |
A61K 039/395 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 20, 2000 |
DE |
100 57 443.2 |
Claims
1. The use of the heavy chain of an antibody, of a functional
derivative, or of a fragment thereof, comprising a CDR3 region
selected from: (a) an amino acid sequence: V L P F D P I S M D V
(I) (b) an amino acid sequence: A L G S W G G W D H Y M D V (II)
(c) an amino acid sequence having a homology of at least 80% with
an amino acid sequence from (a) or (b) (d) an amino acid sequence
having an equivalent ability to bind to GPIIb/IIIa (1) for jointly
inhibiting fibrinogen binding to platelets and vitronectin binding
to endothelial cells (2) for inhibiting angiogenesis and/or (3) for
inhibiting the metastasis of tumors and/or (4) for inhibiting
intimahyperplasia following vascular damage.
2. The use as claimed in claim 1, wherein the heavy chain, the
functional derivative or the fragment thereof furthermore comprises
a CDR1 region selected from: (a) an amino acid sequence: G Y S W R
(III) (b) an amino acid sequence: S Y A M H (IV) (c) an amino acid
sequence which exhibits a homology of at least 80% with an amino
acid sequence from (a) or (b).
3. The use as claimed in claim 1 or 2, wherein the heavy chain, the
functional derivative or the fragment thereof furthermore comprises
a CDR2 region selected from: (a) an amino acid sequence: D I S Y S
G S T K Y K P S L R S (V) (b) an amino acid sequence: V I S Y D G S
N K Y Y A D S V K G tm (VI) (c) an amino acid sequence which
exhibits a homology of at least 80% with an amino acid sequence
from (a) or (b).
4. The use of the light chain of an antibody, of a functional
derivative, or of a fragment thereof, comprising a CDR3 region
selected from: (a) an amino acid sequence: A T W D D G L N G P V
(VII) (b) an amino acid sequence: A A W D D S L N G W V (VIII) (c)
an amino acid sequence which exhibits a homology of at least 80%
with an amino acid sequence from (a) or (b), and (d) an amino acid
sequence having an equivalent ability to bind to GPIIb/IIIa (1) for
jointly inhibiting fibrinogen binding to platelets and vitronectin
binding to endothelial cells (2) for inhibiting angiogenesis and/or
(3) for inhibiting the metastasis of tumors and/or (4) for
inhibiting intimahyperplasia following vascular damage.
5. The use as claimed in claim 4, wherein the light chain, the
functional derivative or the fragment thereof furthermore comprises
a CDR1 region selected from: (a) an amino acid sequence: S G S S S
N I R S N P V S (IX) (b) an amino acid sequence: S G S S S N I G S
N T V N (X) (c) an amino acid sequence having a homology of at
least 80% with an amino acid sequence from (a) or (b).
6. The use as claimed in claim 4 or 5, wherein the light chain, the
functional derivative or the fragment thereof furthermore comprises
a CDR2 region selected from: (a) an amino acid sequence: G S H Q R
P S (XI) (b) an amino acid sequence: S N N Q R P S (XIII) (c) an
amino acid sequence having a homology of at least 80% with an amino
acid sequence from (a) or (b).
7. The use of an antibody, of a functional derivative or of a
fragment thereof, comprising (a) a heavy chain, a functional
derivative or a fragment thereof as in one of claims 1 to 3 (b) a
light chain, a functional derivative or a fragment thereof as in
one of claims 4 to 6 (1) for jointly inhibiting fibrinogen binding
to platelets and vitronectin binding to endothelial cells (2) for
inhibiting angiogenesis and/or (3) for inhibiting the metastasis of
tumors and/or (4) for inhibiting intimahyperplasia following
vascular damage.
8. The use as claimed in one of claims 1 to 7 for the therapy
and/or prophylaxis of vascular occlusion.
9. The use as claimed in one of claims 1 to 7 for tumor
therapy.
10. The use of nucleic acid which encodes a protein as in one of
claims 1 to 9 (1) for jointly inhibiting fibrinogen binding to
platelets and vitronectin binding to endothelial cells (2) for
inhibiting angiogenesis and/or (3) for inhibiting the metastasis of
tumors and/or (4) for inhibiting intimahyperplasia following
vascular damage.
11. The use as claimed in claim 10 for the therapy and/or
prophylaxis of vascular occlusion.
12. The use as claimed in claim 10 for tumor therapy.
Description
DESCRIPTION
[0001] The present invention relates to the use of special,
phage-display-optimized antibodies directed against GPIIb/IIIa for
jointly inhibiting fibrinogen binding to platelets and vitronectin
binding to endothelial cells for the purpose of the therapy and/or
prophylaxis of vascular occlusion. The present invention
furthermore relates to the use of the antibodies for inhibiting
angiogenesis and/or for inhibiting the metastasis of tumors and/or
for inhibiting intima hyperplasia following vascular damage.
[0002] It has been known for a long time that glycoprotein IIb/IIIa
(GPIIb/IIIa, also termed .alpha..sub.IIb.beta..sub.3 or CD41/CD61)
is expressed on the surface of thrombocytes. The receptor is also
frequently termed the fibrinogen receptor since fibrinogen is the
preferred ligand. However, in addition to this, the receptor also
binds a large number of other ligands which contain the RGD
sequence, such as fibronectin, vitronectin and von Willebrand
factor.
[0003] It has also been known for a long time that GPIIb/IIIa plays
an essential role in cellular hemostasis. Normally, thrombocytes do
not remain attached to the vascular endothelium nor do they stick
to each other. However, if the thrombocyte comes into contact with
damaged blood vessels whose endothelium is torn, there then follows
an interaction with the underlying matrix proteins, such as
collagen, fibronectin or laminin, for which the thrombocyte
possesses specific membrane receptors which are similar to the
integrins. However, in blood vessels in which high shearing forces
occur, these interactions are not sufficient for the platelets to
adhere and for a clot to be formed. The latter becomes possible,
inter alia, because the thrombocytes carry the glycoprotein
IIb/IIIa (GPIIb/IIIa) on their surface, which protein mediates
binding to fibrinogen and is therefore also termed the fibrinogen
receptor. Following activation by intracellular messengers, the
GPIIb/IIIa receptor recognizes fibrinogen molecules and in this way
mediates crosslinking of the thrombocytes (Loffler & Petrides,
Biochemie und Pathobiochemie [Biochemistry and Pathobiochemistry],
Springer, Berlin).
[0004] Germ line mutations in the genes for the GPIIb/IIIa receptor
lead to a rare, autosomally recessive bleeding disease which is
characterized by a prolonged bleeding time, normal thrombocyte
values and the complete absence of platelet aggregation and is
termed Glanzmann's thrombasthenia (Loffler & Petrides,
Biochemie und Pathobiochemie, Springer, Berlin).
[0005] Conversely, it is possible to block the GPIIb/IIIa receptor
for the purpose of preventing or treating undesirable vascular
occlusions. Small molecules or else antibodies are suitable for
this purpose. Abciximab (ReoPro) is a human/mouse chimeric
monoclonal antibody Fab fragment which is derived from the murine
monoclonal antibody 7E3 and which binds with great avidity both to
the activated and the non-activated form of GPIIb/IIIa. This
antibody has been licensed as supplementary therapy for preventing
ischemic complications in the heart in patients who undergo a
percutaneous intervention in the coronary blood vessels. Other
indications for abciximab (ReoPro) are unstable angina, stenting in
the carotid, ischemic stroke and peripheral vascular diseases
(Cohen et al., Pathol. Oncol. Res. 6: 163-174 (2000)).
[0006] Cell types which [lacuna] GPIIb/IIIa and
.alpha..sub.5.beta..sub.3 are also thought to be connected with
events in angiogenesis, vascularization and neovascularization.
Hypoxia, as occurs, for example, in diabetes, asthma and
Alzheimer's disease, and inflammatory processes are considered to
be inducers of the angiogenesis. Thereby in many cases, this
angiogenesis is desirable, for example because it counteracts
ischemia following myocardial infarctions and ensures that the
heart tissue is supplied with oxygen and other essential factors.
However, since the blood vessels which are subsequently formed are
frequently not fully differentiated, the subsequent formation of
the blood vessels can also be undesirable. A list of diseases which
are associated with increased vascularization has recently been
published (Carmeliet & Jain, Nature 407 (2000), 249-257).
[0007] In any case, tumor angiogenesis is undesirable. As early as
1971, it was postulated that tumor growth and metastasis proceed in
an angiogenesis-dependent manner (Folkman, J. Cancer Medicine (eds.
Holland, J. F. et al.), 132-152). This made it clear that
inhibiting angiogenesis was a possible strategy for inhibiting
tumor growth and metastasis. Tumor blood vessels differ from normal
blood vessels as a result of their irregular construction, variable
diameter and excessive ramifications and openings between the
endothelial cells delimiting the blood vessel and a discontinuous
or absent basal membrane (Carmeliet & Jain, Nature 407 (2000),
249-257). Although at least some tumor blood vessels possess a
mosaic-like construction composed of endothelial cells and cancer
cells, it remains difficult to specifically recognize
tumor-specific blood vessels. It is true that it has been possible,
by selecting phage-display libraries in vivo, to isolate peptides
which preferentially recognize the blood vessels of subcutaneous
tumors in mice (Arap et al., Science 279 (1998), 377-380). However,
it has still not been demonstrated that these peptides can be used
in vivo to effectively concentrate antineoplastic agents in the
region of the tumor. For this reason, most efforts are still being
directed toward developing angiogenesis inhibitors for the purpose
of treating tumors. A current list of the substances tested in
clinical experiments can be found in Carmeliet (Carmeliet &
Jain, Nature 407 (2000), 249-257).
[0008] However, molecules directed against GPIIb/IIIa are not only
of interest for controlling the growth and metastasis of tumors, on
account of their angiogenesis-inhibiting properties. Thus, it has
become clear, as a result of more recent studies, that expression
of the GPIIb/IIIa receptor is not restricted to thrombocytes. Thus,
Trikha et al. showed, both at the RNA level and the protein level,
that the human. melanoma cell lines WM983B, WM983A and WM35 express
GPIIb/IIIa (Trikha et al., Cancer Res. 57: 2522-2528 (1997)). Timar
et al. have also obtained similar results in the case of B16a
metastatic melanoma cells. These authors also showed that
stimulating cells with a protein kinase C activator stimulates the
translocation of GPIIb/IIIa from an intracellular pool to the cell
surface. The expression of GPIIb/IIIa plays a role in cell adhesion
and, in particular, in tumor cell invasion through the basal
membrane.
[0009] In WO 98/55619, the applicant recently described improved
antibodies directed against GPIIb/IIIa, which antibodies were
optimized by means of phase display and panning selection. The
disclosure in this patent specification contains detailed
information with regard to the hypervariable,
complementarity-determining regions (CDRs) and the framework
regions (FRs).
[0010] The object of the present invention was to search for
further indications for the antibodies described in WO
98/55619.
[0011] This object was achieved by the surprising observation that
the antibodies can be used to jointly inhibit fibrinogen binding to
platelets and vibronectin binding to endothelial cells, which means
that these antibodies are particularly well suited for treating
and/or preventing vascular occlusion. The inhibition of the binding
of vitronectin to endothelial cells may possibly be connected to
the expression of the vitronectin receptor
.alpha..sub.5.beta..sub.3 on endothelial cells. It is assumed that
the antibodies from WO 98/55619 crossreact with the vitronectin
receptor on endothelial cells, and in this way inhibit the binding
of vitronectin, on account of the subunit .beta..sub.3 being
possessed in common. However, it is also possible that the effect
is mediated by other integrin receptors which recognize the RGD
sequence or whose ligand binding can be inhibited by RGD. Because
of the fact that the antibodies crossreact with the vitronectin
receptor .alpha..sub.5.beta..sub.3 on endothelial cells, the
antibodies can also be used for treating intimahyperplasia
following vascular damage. This indication for the WO 98/55619
antibodies follows from the observation that
.alpha..sub.5.beta..sub.3 (=CD51/CD61) is connected with
intimahyperplasia following vascular damage.
[0012] It has furthermore been found, surprisingly, that the
antibodies described in WO 98/55619 possess very promising
properties which make it appear appropriate to use these antibodies
for inhibiting angiogenesis. The antibodies are consequently
suitable for treating tumors and, in particular, for preventing the
metastasis of tumors since they impede the vascularization of the
primary tumor and consequently impede its ability to colonize
surrounding tissue and release proliferating cells into the blood
circulation. In addition, it must be assumed that blocking the
GPIIb/IIIa receptors with the antibodies described in WO 98/55619
impairs the ability of migrated tumor cells to adhere at the target
site and penetrate into the tissue.
[0013] The invention accordingly relates to the use of the heavy
chain of an antibody, of a functional derivative, or of a fragment
thereof, comprising a CDR3 region selected from:
[0014] (a) an amino acid sequence:
V L P F D P I S M D V (I)
[0015] (b) an amino acid sequence:
A L G S W G G W D H Y M D V (II)
[0016] (c) an amino acid sequence having a homology of at least 80%
with an amino acid sequence from (a) or (b)
[0017] (d) an amino acid sequence having an equivalent ability to
bind to GPIIb/IIIa
[0018] for inhibiting angiogenesis and/or for inhibiting the
metastasis of tumors and/or for inhibiting intimahyperplasia
following vascular damage.
[0019] The heavy chain according to the invention, the functional
derivative or the fragment thereof preferably furthermore comprises
a CDR1 region selected from:
[0020] (a) an amino acid sequence:
G Y S W R (III)
[0021] (b) an amino acid sequence:
S Y A M H (IV)
[0022] (c) an amino acid sequence which exhibits a homology of at
least 80% with an amino acid sequence from (a) or (b).
[0023] The heavy chain according to the invention, the functional
derivative or the fragment thereof furthermore preferably comprises
a CDR2 region selected from:
[0024] (a) an amino acid sequence:
D I S Y S G S T K Y K P S L R S (V)
[0025] (b) an amino acid sequence:
V I S Y D G S N K Y Y A D S V K G (VI)
[0026] (c) an amino acid sequence which exhibits a homology of at
least 80% with an amino acid sequence from (a) or (b).
[0027] A further aspect of the present invention is accordingly the
use of the light chain of an antibody, of a functional derivative
or of a fragment, selected from:
[0028] (a) an amino acid sequence:
A T W D D G L N G P V (VII)
[0029] (b) an amino acid sequence:
A A W D D S L N G W V (VIII)
[0030] (c) an amino acid sequence which exhibits a homology of at
least 80% with an amino acid sequence from (a) or (b), and
[0031] (d) an amino acid sequence having an equivalent ability to
bind to GPIIb/IIIa
[0032] for inhibiting angiogenesis and/or for inhibiting the
metastasis of tumors and/or for inhibiting intimahyperplasia
following vascular damage.
[0033] The light chain according to the invention, the functional
derivative or the fragment thereof furthermore preferably comprises
a CDR1 region selected from:
[0034] (a) an amino acid sequence:
S G S S S N I R S N P V S (IX)
[0035] (b) an amino acid sequence:
S G S S S N I G S N T V N (X)
[0036] (c) an amino acid sequence having a homology of at least 80%
with an amino acid sequence from (a) or (b).
[0037] In addition, the light chain according to the invention, the
functional derivative or the fragment thereof preferably
furthermore comprises a CDR2 region selected from:
[0038] (a) an amino acid sequence:
G S H Q R P S (XI)
[0039] (b) an amino acid sequence:
S N N Q R P S (XIII)
[0040] (c) an amino acid sequence having a homology of at least 80%
with an amino acid sequence from (a) or (b)
[0041] Within the meaning of the present invention, the expression
"functional derivative of a chain of a human antibody" is to be
understood as meaning a polypeptide which comprises at least a CDR3
region of the heavy and/or light chain as defined above and,
together with the respective complementary chain of the human
antibody (or a derivative of such a chain), can form an antibody
derivative which possesses a recognition specificity for an antigen
which is equivalent to that of the underivatized antibody.
Preferably, such an antibody derivative has a binding constant of
at least 10.sup.6 l/mol, preferably of at least 10.sup.8 l/mol, for
the given antigen.
[0042] Functional derivatives of chains of a human antibody can be
prepared, for example, by using recombinant DNA techniques to
delete, substitute and/or insert segments of the gene encoding the
given polypeptide.
[0043] Particularly preferred functional derivatives of antibody
chains or antibodies are single-chain antibodies which can be
assembled, for example, from the variable domains of the H chain
and the L chain and, where appropriate, a constant domain. The
preparation of such constructs is described in Hoogenboom et al.,
Immunol. Rev. 130 (1992), 41-68; Barbas III, Methods: Companion
Methods Enzymol. 2 (1991), 119 and Pluckthun, Immunochemistry
(1994), Marcel Dekker Inc., Chapter 9, 210-235.
[0044] Within the meaning of the invention, the expression
"equivalent binding ability" is to be understood as meaning an
identical binding affinity and/or specificity, i.e. epitope
recognition as in the specifically disclosed sequences.
[0045] The present invention also relates to the use of a vector,
which contains at least one copy of a nucleic acid which encodes
one of the above-described antibodies, for inhibiting angiogenesis
and/or for inhibiting the metastasis of tumors and/or for
inhibiting intimahyperplasia following vascular damage. Treating
the patient with nucleic acid instead of protein has a number of
advantages. Whereas storing protein is a relatively elaborate
matter, DNA can be stored without difficulty even over long periods
of time. Another advantage of using DNA is that the antibodies are
correctly processed posttranslationally, for example with regard to
glycosylation.
[0046] Independently of the administration form, use, according to
the invention, of the abovementioned antibodies possesses
substantial advantages as compared with the previously known use of
the ReoPro Fab fragment. These advantages include the fact that the
abovementioned antibodies comprise amino acid sequences which are
entirely of human origin and the danger of an undesirable immune
reaction against the antibodies which are employed is consequently
kept as low as possible.
Sequence CWU 1
1
12 1 11 PRT Artificial Sequence Description of Artificial
SequenceCDR3-region of an optimized antibody against GPIIB/IIIA 1
Val Leu Pro Phe Asp Pro Ile Ser Met Asp Val 1 5 10 2 14 PRT
Artificial Sequence Description of Artificial Sequence CDR3-region
of an optimized antibody against GPIIB/IIIA 2 Ala Leu Gly Ser Trp
Gly Gly Trp Asp His Tyr Met Asp Val 1 5 10 3 5 PRT Artificial
Sequence Description of Artificial Sequence CDR1-region of an
optimized antibody against GPIIB/IIIA 3 Gly Tyr Ser Trp Arg 1 5 4 5
PRT Artificial Sequence Description of Artificial Sequence
CDR1-region of an optimized antibody against GPIIB/IIIA 4 Ser Tyr
Ala Met His 1 5 5 16 PRT Artificial Sequence Description of
Artificial Sequence CDR2-region of an optimized antibody against
GPIIB/IIIA 5 Asp Ile Ser Tyr Ser Gly Ser Thr Lys Tyr Lys Pro Ser
Leu Arg Ser 1 5 10 15 6 17 PRT Artificial Sequence Description of
Artificial Sequence CDR2-region of an optimized antibody against
GPIIB/IIIA 6 Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp
Ser Val Lys 1 5 10 15 Gly 7 11 PRT Artificial Sequence Description
of Artificial Sequence light chain of an optimized antibody against
GPIIB/IIIA 7 Ala Thr Trp Asp Asp Gly Leu Asn Gly Pro Val 1 5 10 8
11 PRT Artificial Sequence Description of Artificial Sequence light
chain of an optimized antibody against GPIIB/IIIA 8 Ala Ala Trp Asp
Asp Ser Leu Asn Gly Trp Val 1 5 10 9 13 PRT Artificial Sequence
Description of Artificial Sequence CDR1-region of an optimized
antibody against GPIIB/IIIA 9 Ser Gly Ser Ser Ser Asn Ile Arg Ser
Asn Pro Val Ser 1 5 10 10 13 PRT Artificial Sequence Description of
Artificial Sequence CDR1-region of an optimized antibody against
GPIIB/IIIA 10 Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Thr Val Asn 1
5 10 11 7 PRT Artificial Sequence Description of Artificial
Sequence CDR2-region of an optimized antibody against GPIIB/IIIA 11
Gly Ser His Gln Arg Pro Ser 1 5 12 7 PRT Artificial Sequence
Description of Artificial Sequence CDR2-region of an optimized
antibody against GPIIB/IIIA 12 Ser Asn Asn Gln Arg Pro Ser 1 5
* * * * *